Target (protein/gene name): serine/ threonine protein phosphatase *NCBI Gene # or RefSeq#:813836 *Protein ID (NP or XP #) or Wolbachia#:3H60 *Organism (including strain):Plasmodium falciparum Etiologic Risk Group (see link below):2 */Disease Information(sort of like the Intro to your MiniResearch Writeup): Plasmodium falciparum is a parasitic protozoan that is involved in Malaria. It is the parasite responsible for roughly 85% of all Malaria cases and the most deadly of the Plasmodium protozoans. P. falciparum is carried by Anopheles mosquitoes and enters the body when a female mosquito injects it into the blood stream while feeding. From there, the P. falciparum travels to the liver via the blood stream, replicates, and then attacks the red blood cells. The parasite is carried through the bloodstream to other organs and can cause a host of symptoms including vomiting, fever, chills, diarrhea, and in severe cases, organ failure.
Serine/threonine protein phosphatase is an important target in P. falciparum. It works in part to regulate mitosis, cell division, apoptosis, and other cell functions, all of which are crucial aspects in the virulence of p. falciparum.
GAAIIFNQDLTFEVQEYMSPSLEVIRETFEENQKLREKVLHCSKIVELEKNVST *Length of your protein in Amino Acids:294 amino acids Molecular Weightof your protein in kiloDaltons using the**Expasy ProtParam**website: 33791.5 MolarExtinction coefficientof your protein at 280 nm wavelength: 24785 TMpred graph Image (http://www.ch.embnet.org/software/TMPRED_form.html). Input your amino acid sequence to it. *CDSGene Sequence(paste as text only):atgagcaaaaaaagcaaagataccaaactgattattctgggcgatgtgcatggccagctgcatgatgtgctgtgg ctgtttaaccgctttggcattccgagcagcaccaacatttatatttttaacggcgatattgcggatcgcggcgaaaacgcgaccgaaatttttattctg ctgtttatttttaaactgagctgcaacgatagcgtgattattaaccgcggcaaccatgaatgcagctatatgaacgaagtgtatggcttttataacgaagtg ctgagcaaatatgataacaccatttttgatctgtttcagaacatttttgaactgctgggcctggcggtgaacgtgcagaaccagatttttgtggtgcatgg cggcctgagccgctatcaggatattaccctgaaagaaattgatgaactggatcgcaaaaaacaggaaattctgcatccggaacagtatgaagata ttgtgatttttgatctgctgtggagcgatccgcagaaaaaagaaggcattggcggcaacgcgcgcggcaacaactgcattacctttggcccggatg tgaccgaaatgtttctgaaaaacaacaacctggatattctgattcgcagccatcaggtgccgaaaaccctgaaaggcattgaaagccatcatgaag gcaaagcattaccctgtttagcgcgagcaactattgcaacaaaattaaaaacctgggcgcggcgattatttttaaccaggatctgacctttgaagtgc aggaatatatgagcccgagcctggaagtgattcgcgaaacctttgaagaaaaccagaaactgcgcgaaaaagtgctgcattgcagcaaaattgtg gaactggaaaaaaacgtgagcacc *GC% Content for gene:44.61%
*NCBI Gene # or RefSeq#: 813836
*Protein ID (NP or XP #) or Wolbachia#: 3H60
*Organism (including strain): Plasmodium falciparum
Etiologic Risk Group (see link below):2
*/Disease Information(sort of like the Intro to your MiniResearch Writeup):
Plasmodium falciparum is a parasitic protozoan that is involved in Malaria. It is the parasite responsible for roughly 85% of all Malaria cases and the most deadly of the Plasmodium protozoans. P. falciparum is carried by Anopheles mosquitoes and enters the body when a female mosquito injects it into the blood stream while feeding. From there, the P. falciparum travels to the liver via the blood stream, replicates, and then attacks the red blood cells. The parasite is carried through the bloodstream to other organs and can cause a host of symptoms including vomiting, fever, chills, diarrhea, and in severe cases, organ failure.
Serine/threonine protein phosphatase is an important target in P. falciparum. It works in part to regulate mitosis, cell division, apoptosis, and other cell functions, all of which are crucial aspects in the virulence of p. falciparum.
http://www.who.int/mediacentre/factsheets/fs094/en/
http://www.parasitesinhumans.org/plasmodium-falciparum-malaria.html
Link to TDR Targets page (if present):http://www.tdrtargets.org/targets/view?gene_id=189
Link to Gene Database page (NCBI, EuPath databases -e.g. TryTryp, PlasmoDB, etc - or PATRIC, etc.) http://plasmodb.org/plasmo/showRecord.do?name=GeneRecordClasses.GeneRecordClass&source_id=PF3D7_1355500&project_id=PlasmoDB
Essentiality of this protein: Significant loss of fitness in bloodstream forms, significant loss of fitness in procyclic forms, significant loss of fitness in differentiation of procyclic forms
Is it a monomer or multimer as biological unit? (make prediction athttp://www.ebi.ac.uk/msd-srv/prot_int/pistart.html):Monomer(3 amino acid chains and 6 ligands) (analysis based on protein in T. gondii)
Complex of proteins?: No
Druggable Target (list number or cite evidence from a paper/database showing druggable in another organism):0.5
*EC#:3.1.3.16
Link to BRENDA EC# page: http://www.brenda-enzymes.org/enzyme.php?ecno=3.1.3.16
-- Show screenshot of BRENDA enzyme mechanism schematic
--Enzyme Assayinformation (spectrophotometric, coupled assay ?, reagents): Continuous or single time point spectrophotometric assay of hydrolysis of p-nitrophenyl phosphate (pNPP)
--Link (or citation) to paper that contains assay information https://books.google.com/books?hl=en&lr=&id=Hz_2HV1GWfMC&oi=fnd&pg=PA153&dq=enzyme+assay+of+serine/threonine+phosphatase&ots=4sZj-96tE4&sig=GLqiXzO1mSK2Jb560Gl5Qp9VuvY#v=onepage&q=enzyme%20assay%20of%20serine%2Fthreonine%20phosphatase&f=false
--Links to assay reagents (substrates) pages. p-nitrophenyl phosphate http://www.sigmaaldrich.com/catalog/product/sigma/p7998?lang=en®ion=US
--List cost and quantity of substrate reagents, supplier, and catalog:
Supplier: Sigma Aldrich
Price/amount: $81.20 for 100mL
Product Number: N2770
Structure (PDB or Homology model)
--PDB # or closest PDB entry if using homology model: 3H60
For Homology Model option:
---- Show pairwise alignment of your BLASTP search in NCBI against the PDB
---- Query Coverage: 80%
---- Max % Identities: 44%
---- % Positives: 62%
---- Chain used for homology: Chain A
Current Inhibitors: cyclosporine A, okadaic acid, trifluoperazine
Expression Information (has it been expressed in bacterial cells): Yes, it has been expressed in E. coli (E. coli M15)
Purification Method: N-terminal affinity tag was added to polypeptide. The His tag was purified through nickel-chelation chromatography. http://www.sciencedirect.com/science/article/pii/S0166685101002602
Image of protein (PyMol with features delineated and shown separately):
*Amino Acid Sequence (paste as text only - not as screenshot or as 'code'):
MSKKSKDTKLIILGDVHGQLHDVLWLFNRFGIPSSTNIYIFNGDIADRGENATEIFILLF
IFKLSCNDSVIINRGNHECSYMNEVYGFYNEVLSKYDNTIFDLFQNIFELLGLAVNVQNQ
IFVVHGGLSRYQDITLKEIDELDRKKQEILHPEQYEDIVIFDLLWSDPQKKEGIGGNARG
NNCITFGPDVTEMFLKNNNLDILIRSHQVPKTLKGIESHHEGKCITLFSASNYCNKIKNL
GAAIIFNQDLTFEVQEYMSPSLEVIRETFEENQKLREKVLHCSKIVELEKNVST
*Length of your protein in Amino Acids: 294 amino acids
Molecular Weightof your protein in kiloDaltons using the**Expasy ProtParam**website: 33791.5
MolarExtinction coefficientof your protein at 280 nm wavelength: 24785
TMpred graph Image (http://www.ch.embnet.org/software/TMPRED_form.html). Input your amino acid sequence to it.
*CDSGene Sequence(paste as text only): atgagcaaaaaaagcaaagataccaaactgattattctgggcgatgtgcatggccagctgcatgatgtgctgtgg
ctgtttaaccgctttggcattccgagcagcaccaacatttatatttttaacggcgatattgcggatcgcggcgaaaacgcgaccgaaatttttattctg
ctgtttatttttaaactgagctgcaacgatagcgtgattattaaccgcggcaaccatgaatgcagctatatgaacgaagtgtatggcttttataacgaagtg
ctgagcaaatatgataacaccatttttgatctgtttcagaacatttttgaactgctgggcctggcggtgaacgtgcagaaccagatttttgtggtgcatgg
cggcctgagccgctatcaggatattaccctgaaagaaattgatgaactggatcgcaaaaaacaggaaattctgcatccggaacagtatgaagata
ttgtgatttttgatctgctgtggagcgatccgcagaaaaaagaaggcattggcggcaacgcgcgcggcaacaactgcattacctttggcccggatg
tgaccgaaatgtttctgaaaaacaacaacctggatattctgattcgcagccatcaggtgccgaaaaccctgaaaggcattgaaagccatcatgaag
gcaaagcattaccctgtttagcgcgagcaactattgcaacaaaattaaaaacctgggcgcggcgattatttttaaccaggatctgacctttgaagtgc
aggaatatatgagcccgagcctggaagtgattcgcgaaacctttgaagaaaaccagaaactgcgcgaaaaagtgctgcattgcagcaaaattgtg
gaactggaaaaaaacgtgagcacc
*GC% Content for gene: 44.61%